title: Domain separation and characterization of PriC, a replication restart primosome factor in Escherichia coli.
abstract: In Escherichia coli the oriC-independent primosome plays an essential role in replication restart after dissociation of the replication DNA-protein complex by DNA damage. Primosome is thought to form via two pathways: one PriA dependent and the other PriA independent. PriC is a key protein in the replication restart of the PriA-independent pathway. In this study, we determined that PriC was divided into two domains. Then, we obtained information that: (i) the C-terminal domain preferentially binds to single-stranded DNA (ssDNA); (ii) the binding of PriC to ssDNA depends on salt concentration; and (iii) the binding site size of PriC is approximately 7-9 nucleotides. The protease digestion of PriC suggested that a possible DNA-binding site is the N-terminus of the C-terminal domain where basic amino acid residues are concentrated. Interestingly, α-helical induction of the C-terminal domain of PriC occurred after the addition of DNAs. Also, we examined the role of heptad repeat of leucine or valine residues in the C-terminal domain and PriC oligomerization. This study describes the structure and function analysis of PriC which forms the primosome complex in replication restart.
authors.affiliation: Department of Protein Structure, Function and Design, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, 812-8582, Japan.
authors.forename: Takahiko
authors.initials: T
authors.lastname: Aramaki
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Abe
authors.forename: Takatoshi
authors.initials: T
authors.lastname: Ohkuri
authors.forename: Tomonori
authors.initials: T
authors.lastname: Mishima
authors.forename: Shoji
authors.initials: S
authors.lastname: Yamashita
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Katayama
authors.forename: Tadashi
authors.initials: T
authors.lastname: Ueda
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D020449:Repetitive Sequences, Amino Acid
chemical_list: D004277:DNA, Single-Stranded; D029968:Escherichia coli Proteins; C069995:PriC protein, E coli